Expression of Cone-Photoreceptor–Specific Antigens in a Cell Line Derived from Retinal Tumors in Transgenic Mice

Tan, Elaine M.L.; Ding, Xi‐Qin; Saadi, Anisse; Agarwal, Neeraj; Naash, Muna I.; Al‐Ubaidi, Muayyad R.

doi:10.1167/iovs.03-1114

Cited by 282 publications

(286 citation statements)

References 27 publications

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“…No 11-cis-retinol was detected in the HPLC analysis, however we identified peak 6 in Figure 1D to be 13-cis retinol due its absorption maxima at 328 nm, which corresponds to the absorption maxima of 13-cis retinol. We have previously shown the absence of RPE65 protein in this cell line, (Tan, Ding, Saadi, Agarwal, Naash and Al-Ubaidi, 2004), The above two independent observations confirm the absence of RPE65 in this cell line. This observation is consistent with recent findings that RPE65 is not expressed in cone photoreceptors (Hemati, Feathers, Chrispell, Reed, Carlson and Thompson, 2005;Seeliger, Grimm, Stahlberg, Friedburg, Jaissle, Zrenner, Guo, Reme, Humphries, Hofmann, Biel, Fariss, Redmond and Wenzel, 2001;Wenzel, von, Oberhauser, Tanimoto, Grimm and Seeliger, 2007).…”

Section: Conversion Of Atr To Atol and Retinyl Ester In 661w Cellssupporting

confidence: 83%

“…The 661W cells were grown in DMEM media (Gibco, Carlsbad, California) supplemented with 10% FBS (Cellgro, Herndon, VA) and 1% antimyocotic-antibiotic (Invitrogen) (Tan, Ding, Saadi, Agarwal, Naash and Al-Ubaidi, 2004). The Müller cell line rMC-1 was grown in DMEM media with 10% FBS, 2mM Glutamine and 1% antimyocotic-antibiotic (Sarthy, Brodjian, Dutt, Kennedy, French and Crabb, 1998).…”

Section: Culture Conditionsmentioning

confidence: 99%

“…For this purpose, we used two cell lines generated from rod dominated retinas: the 661W cell line, derived from a mouse retinal tumor and characterized as a cone specific cell line (Al-Ubaidi, Font, Quiambao, Keener, Liou, Overbeek and Baehr, 1992;Tan, Ding, Saadi, Agarwal, Naash and Al-Ubaidi, 2004), and the Müller cell line rMC-1, a rat derived cell line expressing GFAP and CRALBP, both markers for Müller cells (Sarthy, Brodjian, Dutt, Kennedy, French and Crabb, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Retinoid processing in cone and Müller cell lines

Kanan

Kasus‐Jacobi

Moiseyev

et al. 2008

Experimental Eye Research

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To determine whether cones and Müller cells in the rod dominated retina, cooperate to regenerate the 11-cis retinal chromophore via the retinoid cycle, two cell lines from the rod dominated retinas of Murine were used for this study, 661W a mouse cell line derived from cones and rMC-1, a rat Müller cell line. Retinoid cycle enzymes were analyzed by RT-PCR and their catalytic activity were detected by incubation with retinoids and analyzed by HPLC. We found that, 661W cells are capable of reducing all-trans retinal to all-trans retinol due to the presence of multiple dehydrogenases and generate minor amounts of retinyl-ester. The rMC-1 cells take up all-trans retinol and oxidizes it to all-trans retinal or esterify it to retinyl-ester, but are incapable of isomerizing all-trans retinoids to 11-cis retinoids. This could be a reflection of lack of necessary activities in Müller cells in vivo which suggests that Müller cells do not contribute to retinoid cycling by regenerating 11-cis retinoids. Alternatively, this could be due to the potential that rMC-1, as a transformed cell line, has stopped expressing the proteins needed for the regeneration of 11-cis retinoids.

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Section: Conversion Of Atr To Atol and Retinyl Ester In 661w Cellssupporting

confidence: 83%

Section: Culture Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Retinoid processing in cone and Müller cell lines

Kanan

Kasus‐Jacobi

Moiseyev

et al. 2008

Experimental Eye Research

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“…These recently characterised cells express markers of cone photoreceptors, including blue and green opsins, transducin and X-arrestin, and were derived from a postnatal day 8 mouse retina transformed with the SV40 T-antigen. 30 We demonstrate that following growth factor deprivation, these cells execute an apoptotic death programme that involves parallel activation of calpains and caspases. These results substantiate the existence of more than one death pathway in cone photoreceptors and identify calpains as alternative executioners of cell death.…”

Section: Introductionmentioning

confidence: 83%

“…Retina-derived 661W cells have been characterised as photoreceptor neurons, which express specific markers of cones, the cells responsible for colour perception and bright-light vision. 30,41 They represent an invaluable tool for the study of cone PCD, given the detection problems that can arise in studying dynamic events in cells that account for a mere 2.8% of total photoreceptors in the mouse retina. 28 In spite of the fact that previous studies in our group ruled out the involvement of caspases in several in vitro and in vivo models of photoreceptor degeneration, [42][43][44] (data not shown) and -12 were active during apoptosis of 661W cells.…”

Section: Discussionmentioning

confidence: 99%

Multiple death pathways in retina-derived 661W cells following growth factor deprivation: crosstalk between caspases and calpains

2005

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During development of the mammalian retina, neurons that do not succeed in establishing functional synaptic connections are eliminated by apoptosis, allowing the formation of a finely tuned network. Growth factors play a crucial role in controlling the balance between apoptosis and survival signals not only at developmental stages but also in longterm preservation of retinal functions. In the present work, we explore the apoptotic mechanisms triggered by growth factor deprivation of retina-derived 661W cells. Under serum starvation conditions, these cone photoreceptors underwent cell death with participation of caspase-9, -3 and -12. Interestingly, inhibition of caspases did not prevent apoptosis but only resulted in a temporary delay. We show m-calpain activation in parallel with caspases, indicating that more than one execution pathway is available to cone photoreceptors. Moreover, crosstalk of the caspase and calpain pathways was detected, suggesting a loop that may act to amplify the apoptotic cascade.

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Design of synthetic promoters for controlled expression of therapeutic genes in retinal pigment epithelial cells

Johari

Mercer

Liu

et al. 2021

Biotech & Bioengineering

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Age‐related macular degeneration (AMD) associated with dysfunction of retinal pigment epithelial (RPE) cells is the most common cause of untreatable blindness. To advance gene therapy as a viable treatment for AMD there is a need for technologies that enable controlled, RPE‐specific expression of therapeutic genes. Here we describe design, construction and testing of compact synthetic promoters with a pre‐defined transcriptional activity and RPE cell specificity. Initial comparative informatic analyses of RPE and photoreceptor (PR) cell transcriptomic data identified conserved and overrepresented transcription factor regulatory elements (TFREs, 8–19 bp) specifically associated with transcriptionally active RPE genes. Both RPE‐specific TFREs and those derived from the generically active cytomegalovirus‐immediate early (CMV‐IE) promoter were then screened in vitro to identify sequence elements able to control recombinant gene transcription in model induced pluripotent stem (iPS)‐derived and primary human RPE cells. Two libraries of heterotypic synthetic promoters varying in predicted RPE specificity and transcriptional activity were designed de novo using combinations of up to 20 discrete TFREs in series (323–602 bp) and their transcriptional activity in model RPE cells was compared to that of the endogenous BEST1 promoter (661 bp, plus an engineered derivative) and the highly active generic CMV‐IE promoter (650 bp). Synthetic promoters with a highpredicted specificity, comprised predominantly of endogenous TFREs exhibited a range of activities up to 8‐fold that of the RPE‐specific BEST1 gene promoter. Moreover, albeit at a lower predicted specificity, synthetic promoter transcriptional activity in model RPE cells was enhanced beyond that of the CMV‐IE promoter when viral elements were utilized in combination with endogenous RPE‐specific TFREs, with a reduction in promoter size of 15%. Taken together, while our data reveal an inverse relationship between synthetic promoter activity and cell‐type specificity, cell context‐specific control of recombinant gene transcriptional activity may be achievable.

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Expression of Cone-Photoreceptor–Specific Antigens in a Cell Line Derived from Retinal Tumors in Transgenic Mice

Cited by 282 publications

References 27 publications

Retinoid processing in cone and Müller cell lines

Retinoid processing in cone and Müller cell lines

Multiple death pathways in retina-derived 661W cells following growth factor deprivation: crosstalk between caspases and calpains

Design of synthetic promoters for controlled expression of therapeutic genes in retinal pigment epithelial cells

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